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N9664

Sigma-Aldrich

Polynucleotide Phosphorylase from Escherichia coli

Synonym(s):

Polyribonucleotide: 0rthophosphate Nucleotidyltransferase

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About This Item

CAS Number:
Enzyme Commission number:
MDL number:
UNSPSC Code:
12352204

recombinant

expressed in E. coli

form

solution

enzyme activity

≥250 units/mg protein

mol wt

230 kDa

shipped in

dry ice

storage temp.

−70°C

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General description

Polynucleotide phosphorylase (PNPase) is required for cell viability and mRNA turnover in Escherichia coli.

Application

Polynucleotide phosphorylase (PNP) has been used in a study to show that spontaneous mutations resulting from replication errors are reduced in a PNP-deficient strain. It has also been used in a study to show that the absence of PNPase makes E. coli cells sensitive to UV, which suggests PNP has a role in survival of UV damage.

Biochem/physiol Actions

Polynucleotide phosphorylase from Escherichia coli functions as a exonuclease as well as a poly(A) polymerase and can add C and U nucleotides to poly(A) tails.

Unit Definition

One unit will polymerize 1.0 μmole of ADP releasing 1.0 μmole of inorganic phosphate in 15 minutes, at pH 9.1 at 37 °C.

Physical form

Supplied as a solution in 20 mM Hepes buffer pH 7.9, 0.1 mM EDTA, 2 mM DTT, 12.5 mM MgCl2, 200 mM KCl, 21.4% (w/v) Glycerol

Regulatory Information

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B K Mohanty et al.
Proceedings of the National Academy of Sciences of the United States of America, 97(22), 11966-11971 (2000-10-18)
In vitro, polynucleotide phosphorylase of Escherichia coli can both synthesize RNA by using nucleotide diphosphates as precursors and exonucleolytically degrade RNA in the presence of inorganic phosphate. However, because of the high in vivo concentration of inorganic phosphate in exponentially
Involvement of pnp in survival of UV radiation in Escherichia coli K-12
Rath, D., et al.
Annual Review of Microbiology, 128, 1196-1205 (2012)
H Soreq et al.
The Journal of biological chemistry, 252(19), 6885-6888 (1977-10-10)
A simple procedure for purifying polynucleotide phosphorylase from Escherichia coli cells by means of affinity chromatography on an RNA-Sepharose column is described. The purified enzyme preparation has a specific activity 3500-fold that of the crude extract and is essentially homogeneous
Purification and properties of polynucleotide phosphorylase from Escherichia coli.
Y Kimhi et al.
The Journal of biological chemistry, 243(2), 231-240 (1968-01-25)
I Lisitsky et al.
European journal of biochemistry, 261(2), 468-474 (1999-04-24)
Polyadenylation of mRNA has been shown to target the RNA molecule for rapid exonucleolytic degradation in bacteria. To elucidate the molecular mechanism governing this effect, we determined whether the Escherichia coli exoribonuclease polynucleotide phosphorylase (PNPase) preferably degrades polyadenylated RNA. When

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